Kniss D A, Shubert P J, Zimmerman P D, Landon M B, Gabbe S G
Department of Obstetrics and Gynecology, Ohio State University College of Medicine, Columbus 43210-1228.
J Reprod Med. 1994 Apr;39(4):249-56.
The transport of glucose and amino acids from the maternal to fetal circulation through the placenta is critical to the delivery of fuel for normal fetal growth and development. Little information indicates that transplacental glucose or amino acid transport is influenced by hormones or polypeptide growth factors. We developed a continuous cell line of cytotrophoblastlike cells derived from first-trimester human chorionic villi as a model system to study the regulation of glucose and amino acid transport by insulinlike growth factors (IGFs). Using immunocytochemical and biochemical criteria, the cells were shown to manifest a trophoblastlike phenotype. The cells were maintained in serum-supplemented medium until confluent, at which time they were shifted to serum-free medium for one day. Experiments were initiated by transferring the cells to glucose-free assay buffer and incubating them with IGF-I, IGF-II or insulin. Glucose uptake was measured by the transport of 2-deoxy-D-[1,2-3H]glucose (2[3H]DG) in the presence or absence of cytochalasin B, which has been shown to competitively inhibit glucose uptake. IGF-I, IGF-II and insulin each enhanced 2[3H]DG transport in a dose-dependent fashion. Amino acid transport was measured by incubation of the cells with IGF-I for 60 minutes, followed by a 5-minute challenge with alpha-[methyl-3H]aminoisobutyric acid. IGF-I caused a dose-dependent increase in uptake of the amino acid analog. Radioreceptor assays using [125I]insulinlike growth factor I ([125I]IGF-I) demonstrated that the trophoblast-derived cells contained high-affinity, saturable receptors for IGF-I that also bound IGF-II.(ABSTRACT TRUNCATED AT 250 WORDS)
葡萄糖和氨基酸通过胎盘从母体循环转运至胎儿循环,这对于为胎儿正常生长发育提供营养至关重要。几乎没有信息表明经胎盘的葡萄糖或氨基酸转运受激素或多肽生长因子影响。我们建立了一种源自孕早期人绒毛膜绒毛的细胞滋养层样细胞连续系,作为研究胰岛素样生长因子(IGF)对葡萄糖和氨基酸转运调节作用的模型系统。通过免疫细胞化学和生化标准显示,这些细胞呈现出滋养层样表型。细胞在补充血清的培养基中培养至汇合,此时将其转移至无血清培养基中培养一天。实验通过将细胞转移至无糖测定缓冲液中,并与IGF-I、IGF-II或胰岛素孵育来启动。在有或无细胞松弛素B(已证明其可竞争性抑制葡萄糖摄取)存在的情况下,通过2-脱氧-D-[1,2-3H]葡萄糖(2[3H]DG)的转运来测量葡萄糖摄取。IGF-I、IGF-II和胰岛素均以剂量依赖性方式增强2[3H]DG转运。通过将细胞与IGF-I孵育60分钟,然后用α-[甲基-3H]氨基异丁酸刺激5分钟来测量氨基酸转运。IGF-I导致氨基酸类似物摄取呈剂量依赖性增加。使用[125I]胰岛素样生长因子I([125I]IGF-I)的放射受体分析表明,滋养层来源的细胞含有对IGF-I具有高亲和力、可饱和的受体,该受体也能结合IGF-II。(摘要截断于250字)